基于叶绿体蛋白质组学和代谢组学解析CO2加富缓解黄瓜干旱胁迫的生理机制

项目名称： 基于叶绿体蛋白质组学和代谢组学解析CO2加富缓解黄瓜干旱胁迫的生理机制

项目编号： No.31471918

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 李清明

作者单位： 山东农业大学

项目金额： 82万元

中文摘要： 黄瓜是我国栽培面积最大的蔬菜作物之一，对水分状况非常敏感，干旱胁迫导致黄瓜幼苗叶片膜质过氧化损伤严重，光合电子传递过程受抑。本课题组的前期研究结果表明CO2加富可改善黄瓜的水分状况，提高光合性能和水分利用效率，缓解干旱胁迫引起的负面效应，但对其物质代谢机制缺乏深入了解。本项目拟通过荧光差异双向电泳和MALDI-TOF-TOF/MS技术研究黄瓜叶绿体蛋白差异表达，筛选出干旱胁迫下受CO2加富调控的关键叶绿体蛋白，然后利用生物信息学方法，进行差异蛋白的蛋白功能、编码基因以及亚细胞定位分析。同时利用GC-MS和HPLC-MS技术获得黄瓜叶片代谢谱，筛选出干旱胁迫下黄瓜受CO2调控的代谢物质，然后采用CE和HPLC等技术进一步分析关键候选代谢物的含量，明确受CO2调控且发挥功能的关键代谢物，并对其生物学功能进行验证，为系统阐明CO2加富缓解黄瓜干旱胁迫的分子机制奠定基础，具有重要的理论和现实意义。

中文关键词： 黄瓜；二氧化碳加富；干旱胁迫；蛋白质组学；代谢组学

英文摘要： Cucumber (Cucumis sativus L.) is one of the most popular vegetables cultivated in China, and it is very sensitive to drought stress which can lead to serious injury of lipid peroxidation and inhibition of photosynthetic electron transport of cucumber leaves. Our previous studies showed that CO2 enrichment can meliorate plant water status, enhance its photosynthetic property and water use efficiency, and subsequently alleviate the negative effect of drought stress, but lack of deep understanding about substance metabolism. In the present study, two-dimensional difference gel electrophoresis (2D-DIGE) proteomic approach and MALDI-TOF-TOF/MS are used to study the differential expressed proteins of cucumber chloroplast, select the key protein regulated by CO2 enrichment under drought stress, then analyze the protein function, gene encoding and subcellular localization by bioinformatics methods. Meanwhile, GS-MS and HPLC-MS method are used to analyze the metabolomics profiles of cucumber leaves, and based on this profiles the candidate metabolites regulated by CO2 enrichment under drought stress are preliminarily selected. Then CE and HPLC are used to quantitively analyze the candidate metabolites. At last, the biological function of those metabolites and proteins which really play important role in the alleviation of enriched CO2 on drought stressed cucumber will be validated. Based on the above study, it is expected to make a deeper insight into the physiological mechanism that CO2 enrichment alleviating the adverse effects of drought stress on cucumber.

英文关键词： Cucumber;CO2 enrichment;Drought stress;proteomics;metabolomics